Description
This book details recently developed technologies and conventionally employed cytological proceduresfor the study of X-Chromosome Inactivation. Chapters detail live imaging, bioinformatic methods, fluorescence in situ hybridization, and immunofluorescence, and procedures to optimize the study of molecular mechanism underlying X chromosome inactivation. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, X-Chromosome Inactivation: Methods Protocols aims to be useful for researchers in the field of epigenetics, chromatin, noncoding RNA, and nuclear architecture. 1. Screening for Factors Involved in X Chromosome Inactivation using Haploid ESCs Asun Monfort, Giulio Di Minin, and Anton Wutz 2. Unbiased Genetic Screen to Identify Factors Involved in X-chromosome Inactivation Using a Pooled Bar-coded shRNA Library Benoit Moindrot and Neil Brockdorff 3. Unbiased Genetic Screen to Identify Factors Involved in X-chromosome Inactivation Using a Pooled Bar-coded shRNA Library Benoit Moindrot and Neil Brockdorff 4. Manipulation of Xist Imprinting in Mouse Pre-implantation Embryos Atsushi Fukuda, Akihiro Umezawa, and Hidenori Akutsu 5. Somatic Cell Nuclear Transfer in Mice – Basic Protocol and Its Modification for Correcting X Chromosome Inactivation Status Kimiko Inoue, Shogo Matoba, and Atsuo Ogura 6. Live Imaging of Xist RNA Osamu Masui, Edith Heard, and Haruhiko Koseki 7. Live Imaging of X-chromosome Inactivation and Reactivation Kinetics Shin Kobayashi 8. Visualizing the Dynamics of Inactive X chromosomes in Living Cells Using Antibody-based Fluorescent Probes Yuko Sato, Timothy J Stasevich, and Hiroshi Kimura 9. Visualization of Hidden Epitopes at the Inactive X Chromosome Emma R Stewart and Dawn Coverley 10. Chromosome Spread Analyses of Meiotic Sex Chromosome Inactivation Kris G. Alavattam, Hironori Abe, Akihiko Sakashita, and Satoshi H. Namekawa 11. Simultaneous RNA-DNA FISH in Mouse Pre-implantation Embryos Aristea Magaraki, Agnese Loda, Joost Gribnau, and Willy M. Baarends 12. Combined Immunofluorescence, RNA FISH and DNA FISH in Pre-implantation Mouse Embryos Ikuhiro Okamoto 13. RNA-FISH and Immunofluorescence of Mouse Pre- and Post-implantation Embryos Hirosuke Shiura, Yuka Sakata, Kuniya Abe, and Takashi Sado 14. Experimental Analysis of Imprinted Mouse X-chromosome Inactivation Marissa Cloutier, Clair Harris, Srimonta Gayen, Emily Maclary, and Sundeep Kalantry 15. X-chromosome Inactivation and Escape from X inactivation in Mouse Wenxiu Ma, Giancarlo Bonora, Joel B Berletch, Xinxian Deng, William S. Noble, and Christine M. Disteche 16. Practical Analysis of Hi-C Data: Generating A/B Compartment Profiles Hisashi Miura, Rawin Poonperm, Saori Takahashi, and Ichiro Hiratani
